5-μm sections of FFPE tissue were used for H&E staining, IHC with Ki67 antibody (Biocare Medical), TUNEL (DeadEnd Colorimetric System; Promega), or CD31 (BD Biosciences). Proportions of positively stained cells were counted in three random microscopic fields (400× magnification) in each specimen using ImageJ. Quantification of IHC for Ki67 and TUNEL was performed using the NIH ImageJ plugin IHC Image Analysis Toolbox. Quantification of IHC for CD31 was performed using NIH ImageJ as previously described (Owens et al., 2010 (link)). IHC images for human breast tumors were obtained from THPA (Uhlén et al., 2015 (link)), and images from four representative breast tumors (two ductal and two lobular adenocarcinomas) were examined and analyzed by a collaborating board-certified pathologist with expertise in breast cancer (J. Marotti).
Ki67 antibody
Manufactured by Biocare Medical
The Ki67 antibody is a laboratory reagent used to detect the presence of the Ki67 protein, which is a marker of cell proliferation. The Ki67 protein is expressed during active phases of the cell cycle, making it a useful tool for identifying and quantifying proliferating cells in various biological samples.
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Lab products found in correlation
2 protocols using ki67 antibody
1
Quantitative Histological Analysis of Breast Tumors
2
Immunohistochemistry of Formalin-Fixed Paraffin-Embedded Tissues
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Formalin fixed and paraffin embedded slides were first treated in antigen retrieval buffer at 98C for 15 minutes, and then incubated with primary antibody at 4C overnight or 1 hour at room temperature. The next day, after washing, the samples were incubated with HRP conjugated secondary antibody for 30 minutes. They were then washed and incubated with DAB under close monitoring for color development. The β-Catenin antibody was purchased from BD Transduction Laboratories (Cat# 610153), the Ki-67 antibody was purchased from Biocare Medical (Cat# CRM325), the Cyclin D1 antibody was purchased from Millipore (Cat# 04-1151), the Cyto-keratin antibody was purchased from Dako Cytomation (Cat# Z0622), the HGF antibody was purchased from R&D Systems (Cat# AF-294-NA), and the phosphor-Met antibody was purchased from Cell Signaling (Cat# 3077). Mucicarmine staining kit was purchased from Dako Cytomation (Cat# SL016).
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